scholarly journals Development of a Quantitative Real-Time Detection Assay for Hepatitis B Virus DNA and Comparison with Two Commercial Assays

2000 ◽  
Vol 38 (8) ◽  
pp. 2897-2901 ◽  
Author(s):  
Suzan D. Pas ◽  
Edwin Fries ◽  
Robert A. De Man ◽  
Albert D. M. E. Osterhaus ◽  
Hubert G. M. Niesters
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Real Time ◽  
Dynamic Range ◽  
Detection System ◽  
Hbv Dna ◽  
Control Panel ◽  
Detection Assay ◽  
B Virus ◽  
Real Time Detection

A highly reproducible and sensitive real-time detection assay based on TaqMan technology was developed for the detection of hepatitis B virus (HBV) DNA and compared with two commercially available assays. The assay was validated with the Viral Quality Control panel, which also includes EUROHEP HBV DNA standards. This real-time PCR detection system had a dynamic range of 373 to 1010 genome copies per ml and showed an excellent correlation with both the commercial HBV Digene Hybrid Capture II microplate assay (Digene Diagnostics) and the HBV MONITOR assay (Roche Diagnostics). To demonstrate its clinical utility, four chronically HBV-infected patients treated with lamuvidine were monitored using the three different assays. From the results we concluded that this assay is an excellent alternative for monitoring of HBV-infected patients in routine diagnostics and clinical practice, enabling the analysis of a large dynamic range of HBV DNA in a single, undiluted sample.

scholarly journals Quantitation of Hepatitis B Virus Genomic DNA by Real-Time Detection PCR

1999 ◽  
Vol 37 (9) ◽  
pp. 2899-2903 ◽  
Author(s):  
Aki Abe ◽  
Kazuaki Inoue ◽  
Takeshi Tanaka ◽  
Junko Kato ◽  
Naoki Kajiyama ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Real Time ◽  
Hbv Dna ◽  
Serum Samples ◽  
Standard Curve ◽  
Synthetic Dna ◽  
Hbv Replication ◽  
B Virus ◽  
Real Time Detection

Quantitation of hepatitis B virus (HBV) DNA in serum is a useful method for the monitoring of HBV replication. We attempted to develop a quantitative assay system for HBV DNA that is more sensitive, accurate, and reproducible than existing systems. We detected HBV DNA by real-time detection PCR (RTD-PCR) based on Taq Man chemistry. The efficacy of this assay was evaluated by quantitatively measuring sequential levels of synthetic DNA and DNA in clinical serum samples. The detection limit of this system was as few as 10 DNA copies/reaction. A linear standard curve was obtained between 101 and 108 DNA copies/reaction. The coefficient of variation for both intra- and interexperimental variability indicated remarkable reproducibility. This system detected HBV DNA in 100% of chronic hepatitis B patients tested and never detected HBV DNA in healthy volunteers who were negative for HBV markers. These observations suggest that RTD-PCR is an excellent candidate for a standard HBV quantification method.

Rapid detection and quantitation of hepatitis B virus DNA by real-time PCR using a new fluorescent (FRET) detection system

2004 ◽  
Vol 30 (2) ◽  
pp. 191-195 ◽  
Author(s):  
Sani Hussein Aliyu ◽  
Muktar Hassan Aliyu ◽  
Hamisu M Salihu ◽  
Surendra Parmar ◽  
Hamid Jalal ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Real Time ◽  
Real Time Pcr ◽  
Rapid Detection ◽  
Detection System ◽  
Hepatitis B Virus Dna ◽  
B Virus

Real-time quantitation of hepatitis B virus (HBV) DNA in tumorous and surrounding tissue from patients with hepatocellular carcinoma

2002 ◽  
Vol 68 (4) ◽  
pp. 494-499 ◽  
Author(s):  
Isabella Zanella ◽  
Angelo Rossini ◽  
Daniela Domenighini ◽  
Alberto Albertini ◽  
Elisabetta Cariani
Keyword(s):  
Hepatocellular Carcinoma ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Real Time ◽  
Hbv Dna ◽  
Surrounding Tissue ◽  
B Virus

scholarly journals Real Time PCR HBV-DNA Analysis in HBsAg Positive Patients

2015 ◽  
Vol 10 (2) ◽  
pp. 95-99
Author(s):  
Wasila Rahman ◽  
Muhammad Rabiul Hossain ◽  
Arif Ahmed Khan ◽  
Debashish Saha ◽  
SM Mahbubul Alam ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Antiviral Therapy ◽  
Real Time ◽  
Real Time Pcr ◽  
Armed Forces ◽  
Hbv Dna ◽  
Health Concern ◽  
B Virus ◽  
Pcr Method

Introduction: The hepatitis B virus is a global public health concern and leading cause of chronic liver disease in Bangladesh. For the diagnosis and monitoring of treatment of Hepatitis B virus infection, HBV-DNA detection and quantification is now extensively used worldwide.Objectives: The objective of this study was to detect HBV-DNA by real time PCR method in HBsAg positive patients, to compare the results of HBVDNA detection with HBeAg and Anti-HBe and to monitor the response after antiviral therapy in chronic hepatitis B patients and also to observe the intensity of hepatitis B infection in relation to age and sex.Methods: This was a cross sectional type of study conducted in Armed Forces Institute of Pathology (AFIP), Dhaka Cantonment. In this study, 56 sera of HBsAg positive patients were selected who all were subjected to do HBV-DNA (real time PCR) analysis during the period of 29 July to 30 0ctober, 2013.Results: Out of 56 HBsAg positive patients, HBV-DNA was detected in 34 patients. Among these, 8 (23.5%) patients were HBeAg positive, 16 (47%) patients were anti-HBe positive and 10 (29.5%) were negative for both HBeAg and anti-HBe. Age limit of patients was up to 60 years. HBV-DNA positive patients showed male predominance; 26 (76.5%) patients were male and 8 (23.5%) patients were female. Mean age of the patients was 35±14 years. Among 56 HBsAg positive patients, fifteen were receiving antiviral therapy. Out of them, HBV-DNA was decreased among 4 patients and could not be detected among 11 patients.Conclusion: Real time PCR method of detection of HBV-DNA is very important in patients who are HBeAg negative and this method is also applied to monitor treatment response to antivirals and to detect occult HBV infections immune control phase and also to detect reactivation of HBV cases.Journal of Armed Forces Medical College Bangladesh Vol.10(2) 2014

Signature of chronic hepatitis B virus infection in nails and hair

2020 ◽  
Author(s):  
Haruki Komatsu ◽  
Ayano Inui ◽  
Enkhtaivan Odmaa ◽  
Yoshinori Ito ◽  
Shuichiro Umetsu ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Real Time ◽  
Real Time Pcr ◽  
Hbv Dna ◽  
Chimeric Mice ◽  
Hair Samples ◽  
B Virus ◽  
Chronic Hbv ◽  
Immunochemical Staining

Abstract Background: Hepatitis B virus (HBV) is detected in extrahepatic tissues of individuals with HBV infection. Whether nails and hair contain HBV has been unknown. Methods: We examined two patient groups: those with chronic HBV infection alone (n=71), and those with both chronic HBV and hepatitis delta virus (HDV) infections (n=15). HBV DNA in the patients' fingernails and hair were measured by real-time PCR. Hepatitis B surface antigen (HBsAg) of fingernails was evaluated by an enzyme immunoassay. HDV RNA in fingernails was measured by real-time PCR. Immunochemical staining was performed on nails. We used chimeric mice with humanized livers to evaluate the infectivity of nails.Results: Of the 71 pairs of HBV-alone nail and hair samples, 70 (99%) nail and 60 (85%) hair samples were positive for β-actin DNA. Of those 70 nail samples, 65 (93%) were HBV DNA-positive. Of the 60 hair samples, 49 (82%) were HBV DNA-positive. The serum HBV DNA level of the nail HBV DNA-positive patients was significantly higher than that of the nail HBV DNA-negative patients. The hair HBV DNA-positive patients' serum HBV DNA level was significantly higher compared to the hair HBV DNA-negative patients. The nail HBV DNA level was significantly higher than the hair HBV DNA level. The nails and hair HBV DNA levels were correlated (r=0.325, p<0.05). A phylogenetic tree analysis of the complete genome sequence of HBV isolated from nails and hair identified the infection source. Of the 64 nail samples, 38 (59%) were HBsAg-positive. All 15 pairs of chronic HBV/HDV infection nail and hair samples were β-actin DNA-positive. However, nail HBV DNA was detected in two patients (13%). None of the 15 patients were positive for hair HBV DNA. Nail HDV RNA was detected in three patients (20%). Of the 15 patients, eight (53%) were nail HBsAg-positive. HBsAg and hepatitis delta (HD) antigen were detected in the nails by immunochemical staining. Chimeric mice were not infected with PBS containing HBsAg and HBV DNA elucidated from nails.Conclusions: Nails and hair were the reservoir of HBV DNA. Moreover, nails can contain HBsAg, HDV RNA, and HD antigen.

scholarly journals COBAS AmpliPrep-COBAS TaqMan Hepatitis B Virus (HBV) Test: a Novel Automated Real-Time PCR Assay for Quantification of HBV DNA in Plasma

2007 ◽  
Vol 45 (3) ◽  
pp. 828-834 ◽  
Author(s):  
T. Allice ◽  
F. Cerutti ◽  
F. Pittaluga ◽  
S. Varetto ◽  
S. Gabella ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Real Time ◽  
Real Time Pcr ◽  
Hbv Dna ◽  
Pcr Assay ◽  
Cobas Taqman ◽  
B Virus

scholarly journals Hepatitis B virus DNA in Blood Donors Positive of Anti-Hepatitis B Core Antibodies and Negative for Surface Antigen in Hawler Major Blood Bank, Kurdistan Region, Iraq

2018 ◽  
Vol 60 (1) ◽  
pp. 57-61
Author(s):  
Rasha N. Hassan ◽  
Ali H. Hussain
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Real Time ◽  
Real Time Pcr ◽  
Blood Donors ◽  
Hbv Dna ◽  
Occult Hepatitis ◽  
B Virus ◽  
Occult Hepatitis B ◽  
Hbs Ag

Background: Occult Hepatitis B virus infection (OBI) among blood donors is an important medical concern.Objectives: This study was done to detect the presence of occult hepatitis B virus infections among blood donors with negative hepatitis B surface antigen and positive total anti-hepatitis B core antibodies in Hawler Major Blood Bank in Hawler city/Kurdistan Region of Iraq.Methods: A total number of 12,185 blood donors in Hawler Major Blood Bank were screened for HBsAg and total anti-HBcAb using ELISA technique, and then positive results were retested by confirmatory technique by Chemiluminescence assay. All HBsAg-/HBcAb+ were selected as the study group; HBV DNA was tested among HBsAg-/HBcAb+ by conventional PCR and Real time-PCR. Clinical and demographic data of study group were recorded.Results: Among the 12,185 blood donors, HBsAg was positive in 27 (0.22%) donors using Chemiluminescence assay, the frequency of HBs Ag -/ HBc Ab+ was 276 (2.27%), and then the total prevalence of HBV infection in all blood donors was 2.49%. Among the 276 HBs Ag-/HBcAb+, occult hepatitis B virus infection (OBI) was positive in 39.1% (108/276) using conventional PCR and Real time-PCR techniques, while the prevalence among all blood donors (n=12,185) was 0.09%. Testing of HBV-DNA in HBs Ag -/ HBc Ab+ group for OBI was done by qualitative PCR (positive HBV-DNA=102/276) or by quantitative Real time-PCR (positive HBV-DNA=108/276).Conclusions: The OBI is frequently detected among blood donors in Hawler city especially those have HBsAg-/HBcAb+, and the total anti-HBcAb is an essential serological marker for screening HBV among blood donors. The risk factors for developing OBI among blood donors should be elucidated.   الحامض النووي  الريبوزي منقوص الأكسجين للفايروس الكبدي نوع ب في دم المتبرعين والموجب لأضداد اللب للفايروس وسالب للستضد السطحي للفايروس في مصرف الدم الرئيسي في اربيل/كردستان العراق  رشا نزار حسن علي حاتم حسين نظرة اولية: ان تواجد الاصابة  بالفايروس الكبدي نوع ب لدى المتبرعين بالدم يعتبر من الحالات الطبية التي يجب الانتباه اليها. الاهداف: تم اجراء هذه الدراسة على المتبرعين بالدم في مصرف الدم الرئيسي في مدينة اربيل/كردستان العراق , لغرض التحري عن وجود الاصابات الكبدية الفايروسية نوع ب المخفية لدى المتبرعين بالدم والذين يكون المستضاد السطحي الفايروسي الكبدي نوع ب (HBsAg) سالبا لديهم ويكون المضاد اللبي الكلي لنفس الفايروس(anti-HBcAb) موجبا. طريقة البحث: تم فحص اثنا عشر الفا ومائه وخمس وثمانين متبرع بالدم في مصرف الدم الرئيسي في مدينة اربيل لغرض الكشف عن (HBsAg) و  (anti-HBcAb) وان النتائج الموجبة تم اعادة فحصها بطرق فحص تأكيدية. ان كل المتبرعين ذوو النتائج (السالبة HBsAg ) / و (الموجبة anti-HBcAb) تم انتقئهم كمجموعة الدراسة وتم فحص وجود الحامض النووي  الريبوزي منقوص الأكسجين للفايروس الكبدي نوع ب لديهم في الدم بطريقتين: طريقة تفاعل البوليميراز المتسلسل التقليدية وطريقة تفاعل البوليميراز المتسلسل اللحظي. كما تم تسجيل البيانات الديموغرافية والسريرية لمجموعة الدراسة. النتائج:  لدى فحص المتبرعين البالغ عددهم  اثنا عشر الفا ومائه وخمس وثمانين متبرع تبين وجود سبعة وعشرون اصابة (0.02%)  بالمستضاد (HBsAg) اما عدد المتبرعين الذين ليس لديهم  / HBsAg والموجبة للمضاد anti-HBcAb  فقد كان 276 (2.27%) وقد تبين ان نسبة انتشار الخمج البدي الفايروسي نوع ب لدى المتبرعين كان 2.49%. عند فحص مجموعة الدراسة البالغ عددها 276 والذين كانت نتيجة فحصهم موجبة للمضاد /HBcAb,وسالبة للمستضاد HBsAg ,تبين وجود الخمج الفايروسي الكبدي نوع ب المخفي في 39.1% (108/276),  بينما كانت نسبة انتشار نفس الخمج لدى مجموع المتبرعين الكلي البالغ 12185 هو 0.09%. لقد كانت نتيجة فحص وجود الحامض النووي  الريبوزي منقوص الأكسجين للفايروس الكبدي نوع ب بالطريقة القليدية هي 102 متبرع من مجموعة الدراسة الباغة 276 اما النتيجة لنفس مجموعة الدراسة لنفس الحامض النووي بطريقة تفاعل البوليميراز المتسلسل اللحظي كانت 108. الاستنتاج: لقد بينت هذه الدراسة وجود وجود الخمج الفايروسي الكبدي نوع ب المخفي بصورة متكررة لدى المتبرعين بالدم في مدينة اربيل وخاصة الذين تكون حالتهم  موجبة للمضاد /HBcAb, وسالبة للمستضاد HBsAg كما بينت الدراسة ان فحص  المضاد اللبي للفايروس الكبدي نوع ب هو ضروري عند التحري عن وجود وجود الخمج الفايروسي الكبدي نوع ب لدى المتبرعين بالدم, كما انه من الضروري التقصي عن عوامل الخطورة التي تؤدي الى الاصابة بالخمج الفايروسي الكبدي نوع ب المخفي. الكلمات المفتاحية: المتبرعون بالدم, الخمج الفايروسي الكبدي نوع ب المخفي, مدينة اربيل, مصرف الدم, تفاعل البوليمراز المتسلسل التقليدي واللحظي

scholarly journals Assessment of Hepatitis B Virus DNA Stability in Serum by the Chiron Quantiplex Branched-DNA Assay

1998 ◽  
Vol 36 (2) ◽  
pp. 382-386 ◽  
Author(s):  
Mel Krajden ◽  
Lorraine Comanor ◽  
Oretta Rifkin ◽  
Anna Grigoriew ◽  
James M. Minor ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Dynamic Range ◽  
Linear Regression Analysis ◽  
Hbv Dna ◽  
Dna Integrity ◽  
B Virus ◽  
Preserve Sample ◽  
The Stability ◽  
Statistical Trends

Quantification of hepatitis B virus (HBV) DNA in serum is used to establish eligibility for treatment and to monitor therapeutic response. With the trend toward centralized testing, defining the conditions that preserve sample integrity is of paramount importance. We therefore evaluated the stability of HBV DNA in 26 previously frozen (PF) and 5 fresh, never previously frozen serum specimens. PF specimens, covering a 3-log10 HBV DNA dynamic range, were thawed and stored at −70, 4, 23, 37, and 45°C (±1.5°C) for 0, 24, 72, and 120 h (±2 h) and were refrozen at −70°C prior to testing. Five fresh specimens were split into two groups. Both group FG1 and group FG2 specimens were handled as described above; however, group FG1 specimens were subsequently maintained at 4°C and were never frozen prior to testing. Linear regression analysis of PF specimens demonstrated no significant HBV DNA degradation at ≤4°C over 5 days; however, HBV DNA levels decreased by 1.8, 3.4, and 20% per day at 23, 37, and 45°C, respectively. Three independent statistical methods confirmed that the probability of specimen failure, defined as a loss of 20% or more of HBV DNA and/or coagulation of serum, was lowest at ≤4°C and increased with temperature. Because only 10 to 20% of individual patient specimens demonstrated losses of HBV DNA of ≥20% at 23 or 37°C, sufficient numbers of serum specimens must be evaluated to determine overall statistical trends. In conclusion, HBV DNA integrity in separated serum specimens is preserved for at least 5 days when the specimens are stored at −70 or 4°C.

Development of real-time detection direct test for hepatitis B virus and comparison with two commercial tests using the WHO international standard

2003 ◽  
Vol 18 (11) ◽  
pp. 1264-1271 ◽  
Author(s):  
MOTOKAZU MUKAIDE ◽  
YASUHITO TANAKA ◽  
SATOSHI KATAYOSE ◽  
HIROYUKI TANO ◽  
MITSUHIRO MURATA ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Real Time ◽  
Direct Test ◽  
International Standard ◽  
B Virus ◽  
Real Time Detection
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